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Lucca, Rosemeire Aparecida da Silva de. "Propriedades físico-químicas da lectina KM+ monitoradas por dicroismo circular (CD) e fluorescência. Estimativa do conteúdo de estrutura secundaria por CD". Universidade de São Paulo, 1994. http://www.teses.usp.br/teses/disponiveis/76/76132/tde-02042014-100315/.
Pełny tekst źródłaRecently a new lectin, KM+, isolated from Artocarpus integrifolia seeds was described. KM+ induces neutrophil migration, agglutination of human red blood cells, proliferation of mouse spleen cells and binding with monosacharides D-mannose, D-glicose and α-metil mannoside. This glycoprotein is composed of four monomers, assembled by non covalent bonds, has 500 aminoacids residues/mol, with a Molecular Weight of 52,000 Daltons and 1.8% of carbohydrates [27]. In this work structural changes of KM+ was studied as a function of temperature, pH, chemical denaturing agents as well as the binding with D-mannose. These changes were monitored by circular dichroism (CD) and fluorimetry. Circular Dichroism (CD) spectroscopy was used for the analysis of the secondary structure of KM+ in solution due do its capacity to indicate the presence and to estimate the proportion of α-helix, β-sheet, β-turn and unordered conformations. This measurent can be regarded as a function of the relative orientation of the chromophores responsible for their chiroptical activity. CD spectroscopy is also one of the methods of choice for monitorization of conformational changes in proteins as a function of solvents, pH, temperature, ionic strength and specific or non specific binding. Two programs which are in use for estimation of secondary structure: SSE, using the linear least squares method and CCA, using the simplex method, were evaluated in the present work. SSE uses a set of proteins with known X-ray data as the basis for evaluation while CCA uses only pure proteins experimental CD spectra. Fluorescence spectroscopy is very useful to monitore of protein conformational changes in solution due to the presence of intrinsic fluorophores. Fluorescence Measurements were performed at 25°C. Samples were excited at 280 nm and the emission was monitored in the range 290-450 nm. The maximum emission as a function of pH was at pH 7.0. The wavelength for maximum emission changed from 328 nm at pH 7.0 to 340 nm at pH 10.5. CD spectra were recorded over the range of 185 up to 260 nm. The Secondary structure content estimated by SSE program was: 0% α-helix, 41% β-sheet, 26% β-turn and 32% random with RMS of 12% and CCA program was: 1% α-helix, 35% antiparallel β-sheet, 21% β-turn and/or parallel B-sheet, 28% random, 15% aromatics contributions and dissulfide linkages with RMS of 1%. The fractions of secondary structure obtained when using CCA program were more consistent than those of SSE program. The simulation by CCA program was better probably due to its desconvolution of the spectral contribution of the common secondary structures using experimental CD curves of proteins. The stability of KM+, in PBS, as a function of temperature changes above 55°C but only at 70°C the shape of the CD spectrum is consistent with the loss of the native ordered secondary structure that should accompany protein unfolding. CD spectra of KM+ in water showed conformational changes as a function of temperature was not consistent with denaturated proteins. The unfolding of KM+ by GdnCl and SDS resulted in CD spectroscopic changes: consistent with the increased random structure and disappearance of beta sheet. Using the two denaturing agents together GdnCl and temperature, the denaturation was observed at lower decreased both GdnCl concentration and at lower temperature. The estimation of the number of binding sites for D-mannose was obtained through the fluorescence intensity decrease due to a quenching effect of D-mannose and showed that the stoichiometry of binding was 2 moles of D-mannoseimol of lectin
Silva, Luana Maria Castelo Melo. "Efeito da lectina da alga marinha vermelha Pterocladiella capillace em feridas limpas induzidas em ratos". reponame:Repositório Institucional da UFC, 2012. http://www.repositorio.ufc.br/handle/riufc/18834.
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Based on the need for new formulations that are more efficient and on the properties provided by molecules derived from seaweed, it is believed that these can be effective in healing process. The lectin from the red seaweed Pterocladiella capillacea (PcL) and the polysaccharides of red algae Solieria filiformis (SfP) were initially analyzed in toxicity testing. PcL was applied to the paw edema test followed by measurement of myeloperoxidase (MPO). We evaluated the effect of the seaweed Pterocladiella capillacea lectin (PcL) and algal polysaccharides Solieria filiformis (SfP) in healing wounds in rats induced. Both molecules were submitted to microbiological tests and assayed for the effect on wound healing in wounds clean induced on the back of rats. SfP was used as a possible vehicle for the administration of PcL and compared to Carbopol 940 (C). The gels (0.9%) were analyzed rheological and then applied to the lesions during a treatment period of 10 days, using kollagenase ® as control. The healing process was evaluated on the size of the wounds, levels of MPO and histological analysis. The molecule SfP and PcL is not toxic for the parameters of body weight, organ and biochemical measurements. However, the histological analysis showed minor changes in liver and kidney. PcL (1, 3 and 9 mg / kg, i.v) reduced the edema induced by carrageenan and its inhibitor when administered with mucin was not possible to check the reduction of edema which was confirmed by measurement of MPO. The two molecules were used in microbiological assays and not inhibit growth of any microorganism tested and unable to use SfP as carbon source. The rheological analysis showed that the SfP used in the formulation of the gels (PcL+SfP and SfP) had the characteristic of a pseudoplastic. Macroscopic analysis of wounds showed a reduction in lesion area in the animals treated with PCL, PCL+SfP, PCL+C (53.5 and 60% respectively) on the sixth day of administration. In histological analysis, there was no severe inflammatory infiltrate in the tissues obtained until 4th day of administration of the gels (PcL and PcL+SfP, PcL+C) and Kollagenase® (positive control). On day 6, the untreated animals and those treated only with SfP showed inflammatory infiltrate. The measurement of MPO showed a reduction in the inflammatory process in the samples containing PcL, whose results corroborate the histological analysis. In conclusion, PcL aid in wound repair, suggesting its use as a possible future tool for the treatment of lesions. The biological and pharmacological role of lectins and polysaccharides of seaweed is part of a study area little explored, where a lot of knowledge should be invested since these biomolecules can be promising for the pharmaceutical industry.
Com base na necessidade de obter novas formulações mais eficientes e diante das propriedades apresentadas pelas moléculas oriundas de algas marinhas, acredita-se que estas possam ser eficazes no processo de cicatrização. A lectina da alga marinha vermelha Pterocladiella capillacea (PcL) e os polissacarídeos da alga vermelha Solieria filiformis (SfP) inicialmente foram analisados em ensaio de toxicidade. PcL foi aplicada no ensaio do edema de pata seguido da dosagem de mieloperoxidase (MPO). Avaliou-se o efeito da lectina da alga Pterocladiella capillacea (PcL) e os polissacarídeos das algas Solieria filiformis (SfP) na cicatrização de feridas induzidas em ratos. Ambas as moléculas foram submetidas a ensaios microbiológicos e analisadas quanto ao efeito no processo de cicatrização em feridas limpas induzidas no dorso de ratos. SfP foi utilizado como um possível veículo para a administração de PcL e comparado ao Carbopol 940 (C). Os géis (0,9%) foram submetidos a análise reológica e então aplicados nas lesões durante um período de tratamento de 10 (dez) dias, utilizando kollagenase® como controle. O processo de cicatrização foi avaliado quanto ao tamanho das feridas, dosagem de MPO e análise histológica. PcL e SfP não demonstraram toxicidade quanto aos parâmetros de peso corpóreo, órgãos e dosagens bioquímicas. Entretanto a análise histológica mostrou pequenas alterações no fígado e rim. PcL (1, 3 e 9 mg/kg, i.v.) reduziu o edema induzido por carragenana e quando administrada com seu inibidor mucina não foi possível verificar a redução do edema o qual foi confirmado pela dosagem de MPO. As duas moléculas foram aplicadas em ensaios microbiológicos e não inibiram o crescimento de nenhum micro-organismo testado, os quais também não foram capazes de utilizar SfP como fonte de carbono. A análise reológica mostrou que os SfP utilizados na formulação dos géis (PcL+SfP e SfP) apresentaram a característica de um pseudoplástico. A análise macroscópica das feridas mostrou uma redução da área da lesão nos animais tratados com PcL+SfP e PcL+C (53,5 e 60%, respectivamente) no sexto dia de administração. Na análise histológica, não foi observado infiltrado inflamatório acentuado nos tecidos obtidos até o 4º dia da administração dos géis (PcL+SfP e PcL+C) e Kollagenase® (controle positivo). No 6º dia, os animais não tratados e os tratados apenas com SfP mostraram infiltrado inflamatório. A dosagem de MPO demonstrou redução no processo inflamatório nas amostras contendo PcL, cujo resultado corrobora com a análise histológica. Em conclusão, PcL auxiliou no reparo de feridas, sugerindo seu uso futuro como uma possível ferramenta para o tratamento de lesões. O papel biológico e farmacológico das lectinas e polissacarídeos de algas marinhas faz parte de uma área de estudos ainda pouco explorada, onde muito conhecimento deverá ser investido visto que estas biomoléculas podem ser promissoras para a indústria farmacêutica.
Oliveira, Wilian Rosário de. "Lectinas de Crotalaria spectabilis R.: isolamento, purificação e atividade aglutinante em Leptospira biflexa (saprófita) e L. interrogans (patogênica)". Instituto de Ciências da Saúde, 2014. http://repositorio.ufba.br/ri/handle/ri/23507.
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CAPES
Lectinas são proteínas que se ligam especificamente a resíduos de açúcar e estão envolvidas no processo de reconhecimento celular e sinalização em diversas vias metabólicas. O objetivo deste estudo foi isolar, purificar e investigar a atividade biológica das lectinas de Crotalaria spectabilis R. quanto a sua capacidade de hemaglutinação e aglutinação das linhagens bacterianas: Leptospira biflexa e L. interrogans. Para extração das proteínas, as sementes foram moídas e suas células lisadas em solução NaCl 0,15 M. Após essa etapa, as proteínas foram precipitadas com acetona e sulfato de amônio, dialisadas, liofilizadas e purificadas por cromatografia de filtração e troca iônica. A quantificação proteica foi realizada pelo método de Bradford e o perfil eletroforético obtido por SDS-PAGE. Para testar a atividade biológica das lectinas foram utilizados ensaios de hemaglutinação bem como de aglutinação das linhagens bacterianas. Em nossos resultados, o método de precipitação proteica por acetona resultou em maior extração quando comparado ao método por sulfato de amônio. A lectina de C. spectabilis R. apresentou um peso molecular de 30 kDa e os ensaios de hemaglutinação foram positivos para a proteína. Assim, concluimos que nas sementes de C. spectabilis R. existem lectinas com capacidade de reconhecer receptores presentes na membrana de eritrócitos humanos e promover aglutinação celular. Por fim, as lectinas da planta C. spectabilis R. também foram capazes de aglutinar L. interrogans e L. biflexa, sendo esta reposta mais acentuada na linhagem patogênica.
Lectins are proteins that bind carbohydrate residues with affinity and are involved in the process of cell recognition and signaling in different metabolic pathways. The aim of this study was to isolate, purify and investigate the biological activity of the Crotalaria spectabilis R. lectins in terms of hemagglutination and agglutination capacity of the bacterial strains: Leptospira biflexa and L. interrogans. For protein extraction, the seeds were milled and their cells lysed in 0,15 M NaCl solution. After this step, the proteins were precipitated with acetone and ammonium sulfate, dialyzed, lyophilized and purified by filtration chromatography and ion exchange, respectively. Protein quantification was performed by the Bradford method and the electrophoretic profile by SDS-PAGE. For testing the biological activities of lectins, hemagglutination assays were used as well agglutination of the bacterial strains. In our results, protein precipitation method by acetone resulted in higher yield when compared to ammonium sulfate. The C. spectabilis R. lectin presented a molecular weight of 30 kDa and the hemagglutination assays were positives for the protein. Thus, we conclude that in the C. spectabilis R. seeds there are lectins with capacity to recognize receptors present in the human erythrocytes membrane and to promote cell agglutination. At last, the seed lectin C. spectabilis R. was also able to agglutinate L. interrogans and L. biflexa, this response being stronger in the pathogenic strain.
Bibi, Rashda. "Synthèse de nouveaux dérivés osidiques pour le ciblage de lectines originales". Thesis, Montpellier, Ecole nationale supérieure de chimie, 2012. http://www.theses.fr/2012ENCM0002.
Pełny tekst źródłaRhamnose binding lectines were discovered as new class of lectines in 1990's. Most of these lectines has been isolated from aquatic animals. It was discovered in 1991 that rhamnose specific receptors may be present in human skin while studying the interaction between liposomes incorating synthesitic glycoproteins and keratinocytes. We have synthesized new derivatives of rhamnose to target these lectines
Santos, Adriano dos [UNESP]. "Estudo da afinidade das proteínas rTgMIC1 e rTgMIC4 da Toxoplasma gondii com fetuína e asialofetuína utilizando técnica piezelétrica". Universidade Estadual Paulista (UNESP), 2012. http://hdl.handle.net/11449/92055.
Pełny tekst źródłaCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
As proteínas de micronema TgMIC1 e TgMIC4 (TgMICs) da Toxoplasma gondii fazem parte de um complexo proteico localizado na superfície do parasita responsável pelo processo de adesão e invasão celular. Os objetivos desse trabalho foram estudar dispositivos piezelétricos contendo em cada um, uma das MICs recombinantes (rTgMIC1 ou rTgMIC4) e utilizá‐los na determinação das constantes de afinidade entre elas com a fetuína e asialofetuína, empregando o modelo da Isoterma de Langmuir. Os dispositivos foram desenvolvidos por meio a abordagem de monocamadas automontadas (SAM) mista de tióis, utilizando solução etanólica contendo 2,5 mM de ácido 11‐mercaptoundecanóico (11‐MUA) e 7,5 mM de 6‐mercapto‐1‐hexanol (C6OH). A formação da SAM, realizada em temperatura ambiente por 12 h, foi monitorada pela técnica de Microbalança a Cristal de Quartzo com Fator Dissipativo (QCM‐D) e voltametria cíclica utilizando o par redox [FeII(CN)6]4‐/ FeIII(CN)6]3‐. Os resultados obtidos por ambas as técnicas evidenciaram a formação de SAM rígida e de elevado grau de cobertura superficial após cinética lenta em que processos de adsorção e organização dos tióis ocorreram simultaneamente. Para a imobilização das rTgMICs, solução aquosa contendo 10 mM de EDC (N‐etil‐N‐(dimetilaminopropil) carbodiimida) e 20 mM de NHS (N‐hidroxisuccinimida) foi utilizada para a ativação dos grupos carboxílicos presentes na SAM por 2 h, e o processo acompanhado por QCM‐D apresentou resultados compatíveis com aqueles encontrados na literatura. Pela mesma técnica, foi possível verificar que ambas as rTgMICs se imobilizam sobre o cristal de quartzo após sua incubação com solução 0,15 mg/mL de cada rTgMIC em tampão Tris‐HCl contendo 200 mM de NaCl...
The Toxoplasma gondii micronemal proteins TgMIC1 and TgMIC4 (TgMICs) are part of a protein complex located on the surface of the parasite responsible for the process of cellular adhesion and invasion. The goal of the present work was to study a piezoelectric device containing the recombinant MICs (rTgMIC1 or rTgMIC4) and use it in determining the affinity constants between the MICs with fetuin and asialofetuin, employing the Langmuir isotherm model. The devices were developed using the approach of self‐assembled monolayer (SAM) in ethanolic solution containing 2.5 mM of 11‐mercaptoundecanoic acid (11‐MUA) and 7.5 mM of 6‐mercaptohexanol (C6OH). The SAM formation, held at room temperature for 12 h, was monitored by the Quartz Crystal Microbalance technique with Dissipative Factor (QCM‐D) and cyclic voltammetry using the redox couple [FeII(CN)6]4‐/FeIII(CN) 6]3‐. The results obtained from both techniques showed the formation of a rigid and high surface degree coverage SAM after a slow kinetic process in which adsorption and organization of the thiols occur simultaneously. For the immobilization of rTgMICs, aqueous solution containing 10 mM EDC N‐ethyl‐N‐(dimethylaminopropyl) carbodiimide and 20 mM NHS (N‐hydroxysuccinimide) was used for 2 h, and the process followed by QCM‐D was consistent with those found in the literature. By the same technique it was found that both rTgMICs are immobilized on the quartz crystal after incubation whit solution 0.15 mg/mL of each rTgMIC in Tris‐HCl containing 200 mM NaCl (pH 8.0) for 2 h. Unlike the functionalization of the quartz crystal rTgMIC4, remaining adsorption sites were observed in the process using the rTgMIC1, wherein the blocking step using 0.1% gelatin solution for 2 h was required. Throughout the QCM‐D technique it was possible... (Complete abstract click electronic access below)
Santos, Adriano dos. "Estudo da afinidade das proteínas "rTgMIC1" e "rTgMIC4" da Toxoplasma gondii com fetuína e asialofetuína utilizando técnica piezelétrica /". Araraquara : [s.n.], 2012. http://hdl.handle.net/11449/92055.
Pełny tekst źródłaBanca: Emanuel Carrilho
Banca: Maria Cristina Roque Antunes Barreira
Resumo: As proteínas de micronema TgMIC1 e TgMIC4 (TgMICs) da Toxoplasma gondii fazem parte de um complexo proteico localizado na superfície do parasita responsável pelo processo de adesão e invasão celular. Os objetivos desse trabalho foram estudar dispositivos piezelétricos contendo em cada um, uma das MICs recombinantes (rTgMIC1 ou rTgMIC4) e utilizá‐los na determinação das constantes de afinidade entre elas com a fetuína e asialofetuína, empregando o modelo da Isoterma de Langmuir. Os dispositivos foram desenvolvidos por meio a abordagem de monocamadas automontadas (SAM) mista de tióis, utilizando solução etanólica contendo 2,5 mM de ácido 11‐mercaptoundecanóico (11‐MUA) e 7,5 mM de 6‐mercapto‐1‐hexanol (C6OH). A formação da SAM, realizada em temperatura ambiente por 12 h, foi monitorada pela técnica de Microbalança a Cristal de Quartzo com Fator Dissipativo (QCM‐D) e voltametria cíclica utilizando o par redox [FeII(CN)6]4‐/ FeIII(CN)6]3‐. Os resultados obtidos por ambas as técnicas evidenciaram a formação de SAM rígida e de elevado grau de cobertura superficial após cinética lenta em que processos de adsorção e organização dos tióis ocorreram simultaneamente. Para a imobilização das rTgMICs, solução aquosa contendo 10 mM de EDC (N‐etil‐N‐(dimetilaminopropil) carbodiimida) e 20 mM de NHS (N‐hidroxisuccinimida) foi utilizada para a ativação dos grupos carboxílicos presentes na SAM por 2 h, e o processo acompanhado por QCM‐D apresentou resultados compatíveis com aqueles encontrados na literatura. Pela mesma técnica, foi possível verificar que ambas as rTgMICs se imobilizam sobre o cristal de quartzo após sua incubação com solução 0,15 mg/mL de cada rTgMIC em tampão Tris‐HCl contendo 200 mM de NaCl... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: The Toxoplasma gondii micronemal proteins TgMIC1 and TgMIC4 (TgMICs) are part of a protein complex located on the surface of the parasite responsible for the process of cellular adhesion and invasion. The goal of the present work was to study a piezoelectric device containing the recombinant MICs (rTgMIC1 or rTgMIC4) and use it in determining the affinity constants between the MICs with fetuin and asialofetuin, employing the Langmuir isotherm model. The devices were developed using the approach of self‐assembled monolayer (SAM) in ethanolic solution containing 2.5 mM of 11‐mercaptoundecanoic acid (11‐MUA) and 7.5 mM of 6‐mercaptohexanol (C6OH). The SAM formation, held at room temperature for 12 h, was monitored by the Quartz Crystal Microbalance technique with Dissipative Factor (QCM‐D) and cyclic voltammetry using the redox couple [FeII(CN)6]4‐/FeIII(CN) 6]3‐. The results obtained from both techniques showed the formation of a rigid and high surface degree coverage SAM after a slow kinetic process in which adsorption and organization of the thiols occur simultaneously. For the immobilization of rTgMICs, aqueous solution containing 10 mM EDC N‐ethyl‐N‐(dimethylaminopropyl) carbodiimide and 20 mM NHS (N‐hydroxysuccinimide) was used for 2 h, and the process followed by QCM‐D was consistent with those found in the literature. By the same technique it was found that both rTgMICs are immobilized on the quartz crystal after incubation whit solution 0.15 mg/mL of each rTgMIC in Tris‐HCl containing 200 mM NaCl (pH 8.0) for 2 h. Unlike the functionalization of the quartz crystal rTgMIC4, remaining adsorption sites were observed in the process using the rTgMIC1, wherein the blocking step using 0.1% gelatin solution for 2 h was required. Throughout the QCM‐D technique it was possible... (Complete abstract click electronic access below)
Mestre
Destecroix, Harry. "Anthracene based synthetic lectins". Thesis, University of Bristol, 2015. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.682557.
Pełny tekst źródłaAndersson, Pontus. "Comparison of Lectins and their suitability in Lectin Affinity Chromatography for isolation of Glycoproteins". Thesis, Uppsala universitet, Institutionen för biologisk grundutbildning, 2020. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-417024.
Pełny tekst źródłaSousa, Bruno Lopes de. "CaracterizaÃÃo estrutural das formas silvestre e recombinante de uma lectina de sementes de Vatairea macrocarpa Benth e anÃlise das suas bases moleculares de ligaÃÃo ao antÃgeno Tn". Universidade Federal do CearÃ, 2014. http://www.teses.ufc.br/tde_busca/arquivo.php?codArquivo=13196.
Pełny tekst źródłaAs lectinas consitem em uma classe diversificada de proteÃnas, capazes de reconhecer estruturas glicÃdicas de forma reversÃvel e com alta especificidade, no entanto sem alterar suas estruturas quÃmicas, participando de vÃrios processos celulares importantes. Dentre as diferentes famÃlias de lectinas, as isoladas a partir de leguminosas sÃo as mais extensivamente estudadas, havendo sido relatada a influÃncias dessas molÃculas sobre diversos processos patolÃgicos, incluindo a carcinogÃnese. NotÃveis propriedades antitumorais tÃm sido detectadas para algumas lectinas de leguminosas, resultantes da sua habilidade em induzir a morte celular ou a autofagia em cÃlulas cancerÃgenas, o que atraÃdo atenÃÃo para suas possiveis aplicaÃÃes biomÃdicas. AlÃm disso, algumas lectinas desse grupo especificas para galactose/N-acetil-D-galactosamina (Gal/GalNAc) tÃm se mostrado Ãteis como marcadores histoquÃmicos na pesquisa do cÃncer e a caracterizaÃÃo estrutural dessas lectinas em complexo com diferentes epÃtopos cancerÃgenos vem sendo realizada com sucesso. A lectina isolada a partir das sementes da leguminosa Vatairea macrocarpa (VML) à uma lectina bem caracterizada especÃfica para Gal/GalNAc capaz de reconhecer especificamente o antÃgeno Tn (GalNAc-α-O-Ser), naturalmente encontrado em O-mucinas presentes em diferentes tipos de cÃncer. As estruturas cristalogrÃficas para a VML em complexo com o antÃgeno Tn e GalNAc foram determinadas com resoluÃÃes de 1.4 e 1.7 Ã, respectivamente. A maioria das lectinas obtidas a partir de fontes naturais consiste em misturas de diferentes isoformas, uma caracterÃstica indesejada para aplicaÃÃes biomÃdicas. Com base nisso, uma construÃÃo recombinante para VML (rVML) foi expressa em Escherichia coli, sendo obtida de forma solÃvel em com alto rendimento. A estrutura cristalina para a rVML, bem como para seus complexos com o antÃgeno Tn, GalNAc e α-Lactose foram determinadas com resoluÃÃes de 1.7, 2.7, 2 e 1.8 Ã, respectivamente, apresentando a mesma estrutura geral e padrÃes de interaÃÃo que a lectina silvestre. Com o intuito de gerar um perfil comparativo entre a VML e outras lectinas de leguminosas capazes de reconhecer o antÃgeno Tn, foram realizadas anÃlises de docking molecular utilizando fragmentos de O-mucinas diferentemente decorados com o antÃgeno Tn. Esse perfil ressalta como alteraÃÃes sutis no elenco ou disposiÃÃo dos aminoÃcidos constituintes do sÃtio de ligaÃÃo a carboidrato, que talvez nÃo influenciem a capacidade de ligaÃÃo a monossacarÃdeo, podem impactar diretamente a habilidade dessas lectinas em reconhecer antÃgenos em condiÃÃes naturais. Adicionalmente aos jà caracterizados efeitos biolÃgicos relatados para VML, a similaridade entre sua estrutura e perfis de interaÃÃes quando comparadas a outras lectinas comumente utilizadas como marcadores histoquÃmicos (e.g., VVLB4 e SBA), sugerem fortemente a possÃvel utilizaÃÃo da VML como uma nova ferramenta na pesquisa do cÃncer. Esse trabalho consiste no primeiro relato de estruturas cristalogrÃficas para uma lectina de leguminosa especÃfica para Gal/GalNAc da tribo Dalbergieae.
Lectins are a very diverse class of proteins able to bind specific sugar structures reversibly and with high specificity, but without enzymatically modifying them, triggering several important cellular processes. Among the different lectin families, legume lectins are the most thoroughly studied and have been widely reported to exhibit a number of links to many pathological processes, including carcinogenesis. The remarkable anti-tumor properties of some legume lectins, resulting from their ability to induce programmed cell death and/or autophagocytosis in cancer cells have attracted much attention for biomedical applications. Moreover, a few galactose/N-acetylgalactosamine (Gal/GalNAc)-binding lectins from this group have proven to be useful markers for cancer histochemistry, and the structural characterization of these lectins bound to specific cancer epitopes has been carried out successfully. The seed lectin isolated from the legume tree Vatairea macrocarpa (VML) is a well characterized Gal/GalNAc-binding protein able to specifically recognize naturally occurring O-mucins presenting the carcinoma epitope Tn antigen (GalNAc-α-O-Ser). The crystal structures of VML in complex with Tn antigen and GalNAc have been determined at the resolution of 1.4 and 1.7 Ã, respectively. Unfortunately, most of lectins obtained from natural sources consist in a mixture of forms, which is an undesired feature for biomedical applications. Thus, the recombinant form of VML (rVML) was expressed in Escherichia coli, being obtained soluble and wih high yielding. The crystal structure for rVML, as well as for the complex with Tn antigen, GalNAc and α-Lactose have been determined at resolutions of 1.7, 2.7, 2 and 1.8 Ã, respectively, presenting the same overall structure and binding patterns as the wild lectin. Molecular docking analysis of this new structure and other Tn-binding legume lectins to O-mucin fragments differently decorated with this antigen provides a comparative binding profile among these proteins, stressing that subtle alterations that may not influence monosaccharide binding can, nonetheless, directly impact the ability of these lectins to recognize naturally occurring antigens. In addition to the specific biological effects of VML, the structural and binding similarities between it and other lectins commonly used as histological markers (e.g., VVLB4 and SBA) strongly suggest that VML can be used as a new tool for cancer research. This is the first report of crystal structures of a Gal/GalNAc-binding legume lectin from the Dalbergieae tribe.
Howgego, Joshua David. "Synthetic lectins with novel selectivity". Thesis, University of Bristol, 2012. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.573385.
Pełny tekst źródłaPeron, Gabriela. "Tabhys: um peptídeo com atividade lectínica extraído de Tabernaemontana hystrix". Universidade de São Paulo, 2015. http://www.teses.usp.br/teses/disponiveis/17/17147/tde-11112015-155331/.
Pełny tekst źródłaLectins are proteins that have at least one non-catalytic domain that binds specifically and reversibly to a monosaccharide or oligosaccharide. This ability to bind to different types of sugars makes these molecules useful tools in the study of various specific cellular processes. Although the plant lectins are widely studied, those belong to Apocynaceae family are still little explored. Previous results obtained by our research group showed that bark crude extracts from Tabernaemontana hystrix Steud (Apocynaceae) had hemagglutination activity. Besides to agglutinate erythrocytes from ABO blood group system, the putative agglutinin induced the synthesis of IL-6 and TGF-beta mRNA in mouse spleen cells. Here we aim to identify, characterize biochemically and evaluate the possible immunostimulatory potential of T. hystrix agglutinin. The haemagglutination activity was obtained from crude extracts of bark flour, but not of flours of stems without bark and leaves. The activity of the bark extract was similar to that from the previous study, since the haemagglutination was inhibited by the glycoprotein fetuin, but not by monosaccharides. An isolation protocol was determined by using ammonium sulfate precipitation, with haemagglutination activity recovered in the range of 30-60% of saturation, and sequential chromatography procedures: (1) hydrophobic interaction (HiTrap Octyl), (2) cation-exchange (HiTrap SP), (3) reverse phase (EC Nucleosil) and (4) affinity (BlueSepharose) chromatography. From these columns the activity was recovered in the (1) unbound material, and eluates (2 and 4) with 1M and 0,5M of NaCl, respectively, and (3) 83% acetonitrile. On the basis of electrophoresis analysis, the protocol produced a preparation comprised of only band corresponding a peptide with molecular weight (MW) of about 3-kDa, with minimum haemagglutination concentration of 50g/ml. To determine if this molecule arrangement had a quaternary structure arrangement, a feature of most known lectins, we submitted the preparation to a native electrophoresis. Because there was neither change in migration pattern nor presence of molecules of higher molecular mass, we suggested that T. hystrix peptide (Tabhys) is a peptide with MW of about 3-kDa. Since hevein, which is a most studied lectin-like peptide with antifungal activity, binds specifically to chitin, we performed an affinity chromatography in the chitin column with bark extract. We observed haemagglutination activity and the presence of peptide with MW of 3-kDa in the material bound to column and eluted with 0,1M acetic acid. Curiously, this peptide was not able to inhibit the growth of the fungus Trichophyton rubrum. Thereafter, when the purified peptide was used to stimulate murine spleen cells, we detected the expression of TGF-beta message, with a peak production obtained in cell stimulated with 37 ng/mL of Tabhys. In the current study, we isolated a peptide from crude extract of T. hystrix bark with haemagglutination activity, providing new perspectives in molecular and structural researches of peptide lectins.
Didak, Blanka. "Synthèse de néoglycoconjugués et dendrimères glycomimétiques utilisés pour le développement de puces à lectines de type C et leur validation". Thesis, Université Grenoble Alpes (ComUE), 2018. http://www.theses.fr/2018GREAV036.
Pełny tekst źródłaC-type lectins (CLRs) are glycan binding proteins which recognize sugars in Ca2+ dependent manner. They have diverse roles in human organism. They are responsible for interactions and internalization of pathogens like Candida albicans, Mycobacterium tuberculosis, HIV or Ebola virus. They are also involved in development or prevention of cancer through recognition of specific glycans expressed on surface of tumor cells.The crucial importance is to find ligands for CLRs which will induce immune response or inhibitors for lectins which are involved in promotion of infections in organism. The goal of IMMUNOSHAPE network was to closely examine roles, specificities and differences between CLRs and try to find appropriate molecules for driving immune response. During this thesis, neoglycoproteins (NGPs) containing glycodendrons with different valences were synthetized. For synthesis were used BSA and OVA as protein carriers on which are coupled monovalent compounds and dendrons with αMan and Manα1-2Man in three and nine valences. All NGPs were synthetized with click chemistry with two ratios of glycodendron/BSA. Additionally, NGPs with glycomimetics on the basis of fucose and Manα1-2Man were prepared.The affinity of synthetized molecules was analyzed with GLYcoPROFILE, technology platform developed in GLYcoDiag with the aim of better and more precise investigation of glycobiological interactions. It was evaluated that all tested compounds showed strong multivalent effect on four mannose specific lectins, including two CLRs: DC-SIGN and Langerin. Neoglycoprotein with 11 nonavalent Manα1-2Man dendrons achieved the best avidity for all tested lectins. Obtained IC50 for Langerin and DC-SIGN are of nanomolar and picomolar range respectively, which are one of the lowest values obtained for these two lectins.Studying glycobiological interactions was expanded by analysis of differences between glycans with O-, C- and S-linkage and their interactions with lectins. It was observed that C-glycans does not have the same mode of binding like O-glycans and subsequently, do not show the same expected multivalent effect such as O-glycans. In the context of glycosides, we show that even O-glucoside achieved slithly better interactions with purified lectins in comparison with S-glucoside, S-galactoside provide significantly better inhibition between normal human keratinocytes and corresponding neoglycoproteins in comparison with O-galactoside. Furthermore, interesting study was investigation of interactions between thio-sialosides and sialidase NanA. This analysis showed that multivalency has a strong effect not only on lectin, but also on enzyme binding. Synthetized thio-sialylated NGPs showed as efficient, multivalent inhibitor of NanA enzyme.Altogether, this thesis presents interesting results of significant influence of multivalent compounds on glycobiological interactions between glycans and lectins as well as enzymes. This knowledge could be used in future design of vaccines and their adjuvants for infection and cancer treatment and in general, for shaping immune response
Murarasu, Thomas. "The Shiga Toxin B-Subunit : a Promising Scaffold for the Targeting of Tumor Specific Glycosphingolipids". Thesis, Université Paris-Saclay (ComUE), 2017. http://www.theses.fr/2017SACLS512.
Pełny tekst źródłaCancer is the second cause of death worldwide. Recent advance in cancer treatments involved the identification of cancer biomarkers and the development of efficient therapeutic products able to specifically recognize them. This new class of products has the ability to specifically target tumor cells, with the major advantages to decrease or abolish treatments side effects and relapses of the disease. Unfortunately, a certain number of patients do not respond to those treatments lacking the expression of those biomarkers on their tumor. This project aims at developing new targeted therapies by exploiting a new class of cancer biomarkers, which would potentially extend the therapeutics options against cancer
Coelho, Mirela Batista. "Estudo da atividade inseticida e pro-inflamatoria da lectina isolada de sementes de 'Annona coriacea' Mart". [s.n.], 2006. http://repositorio.unicamp.br/jspui/handle/REPOSIP/314522.
Pełny tekst źródłaTese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia
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Resumo: As lectinas são um grupo de proteínas e/ou glicoproteínas que se ligam específica e reversivelmente a carboidratos. Estudos têm demonstrado que essas proteínas possuem importantes atividades biológicas como atividade inseticida contra insetos e respostas imunológicas e fisiológicas em animais. Neste trabalho, tivemos como objetivos comparar os efeitos da lectina de sementes de Annona coriacea (ACLEC), com massa molecular de aproximadamente 14 kDa, sobre o desenvolvimento dos lepidopteras Anagasta kuehniella e Corcyra cephalonica assim como o estudo dos índices nutricionais desses insetos, e investigar a habilidade dessa lectina em induzir a migração de leucócitos em camundongos e os mecanismos farmacológicos delineadores desse processo, utilizando peritonite como modelo experimental. Os resultados da atividade inseticida de ACLEC mostraram que a lectina não produziu efeitos significativos sobre a sobrevivência e o peso larval de Corcyra cephalonica alimentada em dieta contendo até 2,0% de ACLEC, porém, para Anagasta kuehniella, em uma dieta artificial contendo 1,5% e 1,0% de ACLEC produziu um LD50 e WD50, respectivamente. Os resultados dos experimentos do consumo alimentar dos insetos apresentaram apenas uma redução na eficiência de conversão do alimento ingerido (ECI) e do alimento digerido (ECD) em larvas de A. kuehniela. A presença de ACLEC (2,0%) na dieta alimentar em ambos os insetos não modificou a digestibilidade aproximada (AD), mas promoveu um aumento no custo metabólico (CM) e um aumento da atividade proteolítica nas fezes do lepidóptero A. kuehniella. Este mecanismo pode promover uma modificação no meio ambiente da membrana e consequente disrupção do mecanismo de reciclagem ede enzimas, idicando a possibilidade de utulizar esta lectinas como uma estratégia biotecnológica para o manejo de insetos. Para a análise da atividade proinflamatória de ACLEC, camundongos Swiss machos foram injetados intraperitonealmente com ACLEC (3-100 µg/cavidade), de 4 a 96 h e logo após a contagem de leucócitos no fuído do lavado peritoneal foi avaliado. ACLEC induziu uma acumulação de neutrófilos dose-dependente, alcançando a respsota máxima a 16 h após a injeção (aproximadamente um aumento de 40x para 30 µg/cavidade). Uma significante acumulação de células mononucleares foi observada a 72 h (aumento de 2,7x). O carboidrato manose aboliu o inlfuxo de neutrófilos, porém sacarose e glicose não tiveram efeito. Dexametasona, o inibidor de ciclooxigenase-2 celecoxibe e o antagonista do receptor PAF PCA4248 significantemente reduziram o influxo de neutrófilos induzido por ACLEC. O antagonista de taquicinina NK1 SR140333, o antagonista de taquicinina NK2 SR48968, o inibidor não seletivo de NO L-NAME, o inibidor seletivo de NOS Aminoguanidina e o inibidor de lipoxigenase AA861 todos falharam em modificar a resposta induzida por ACLEC. Em conclusão, ACLEC é capaz de atrair neutrófilos na cavidade peritoneal de camundongos por mecanismos envolvendo interações da lectinas com o reconhecimento específico de resíduos de manose presentes nas células, induzindo a liberação de mediadores derivados de COX-2 e PAF
Abstract: Lectins are a group of proteins and/or glycoproteins, which exhibit specific and reversible carbohydrate-binding activities. Studies have demonstrated that such proteins possess important biological activities including insecticide activity as well as immunological and physiological responses in animals. In this investigation, our aim was to compare the effects of lectin isolated of Annona coriacea (ACLEC) seeds, with a molecular mass of 14 kDa, on the development of Anagasta kuehiella and Corcyra cephalonica Lepidopteras, as well as the study of their nutritional index. Since plant lectins are known to present inflammatory activity, this study also sought to investigate the leukocyte migration induced by ACLEC, and inflammatory mediators involved in this phenomenon. The results of insecticide activity of ACLEC showed that the lectin did not produce significant effects on survival and weight of C. cephalonica on an artificial diet of 2,0% of ACLEC, however, for A. kuehniella,on an artificial diet containing 1.5% and 1.0% ACLEC, it produced a LD50 and WD50, respectively. The results from dietary utilization experiments carried out with caterpillars presented only a reduction in efficiency of conversion of ingested food (ECI) and digested food (ECD) in A. kuehniella. ACLEC in the diet did not modify the approximate digestibility (AD) of any insect, but an increase of CM (metabolic cost) and an increase of proteolytic activity in the faeces were observed in A. kuehniella. These results indicate that ACLEC possesses an insecticide effect only towards A. kuehniella larvae, possibly through the binding of lectin on chitin components of membrane peritrophic or glycosylated proteins in the insect midgut. This mechanism can promote a change in membrane environment with the consequent disruption of enzyme recycling mechanisms, indicating the possibility of using this lectin in a biotechnological strategy for insect management. To test proinflammatory activity of ACLEC, male Swiss mice were intraperitoneally injected with ACLEC (3-100 µg/cavity), and at 4 to 96 h thereafter the leukocyte counts in peritoneal washing fluid were evaluated. ACLEC induced a dose-dependent neutrophil accumulation, reaching maximal responses at 16 h after injection (approximately 40-fold increase for 30 µg/cavity). Significant accumulation of mononuclear cells was observed at 72 h (2.7-fold increase). The carbohydrate mannose nearly abolished the neutrophil influx, whereas sucrose and glucose had no effect. Dexamethasone, the cyclooxygenase-2 inhibitor celecoxib and the PAF receptor antagonist PCA4248 significantly reduced ACLEC-induced neutrophil influx. The tachykinin NK1 antagonist SR140333, the tachykinin NK2 antagonist SR48968, the non-selective NO inhibitor L-NAME, the selective inducible NOS inhibitor aminoguanidine and the lypoxygenase inhibitor AA861 all failed to modify the ACLEC-induced responses. In conclusion, ACLEC is able to attract neutrophils into the mice peritoneal cavity by mechanisms involving interactions of the lectin with cell-specific mannose recognition, leading to the release of COX-2-derived mediators and PAF
Doutorado
Bioquimica
Doutor em Biologia Funcional e Molecular
Goto, Leandro Seiji. ""Estudos estruturais e funcionais sobre duas lectinas: cadeia B recombinante da pulchellina & Camptosemina"". Universidade de São Paulo, 2007. http://www.teses.usp.br/teses/disponiveis/76/76132/tde-29032007-121740/.
Pełny tekst źródłaLectins have been placed in a structurally diverse group of proteins that bind carbohydrates and glycoconjugates with high specificity. Found in several organisms, they are extremely useful molecules in the characterization of saccharides, as drug delivery mediators, and even as cellular surface markers. Pulchellin is a D-Galactose-binding heterodimeric glycoprotein from Abrus pulchellus seeds and classified as a type-2 ribosome-inactivating protein (type-2 RIP). The recombinant pulchellin B (rPBC) was previously produced in E. coli BL21 (DE3). In the present work, rPBC is analyzed with respect to its interaction with mammal cells in vitro and structural stability. Results show that rPBC has selectivity for targeted cell types and that it is actively endocytosed, probably by the same route described for other RIPs. Furthermore, rPBC was bond to the catalytic chain and the resulting heterodimer has shown toxicity degree very similar to the native holotoxin, confirming rPBC activity and attributing it a essential role in the intoxication mechanism. A second part of this work has dealed with another lectin extracted from the seeds of Camptosema ellipticum. Recently, experiments with water-soluble extracts obtained from the seed of Camptosema ellipticum have shown to have hemagglutinative properties when assayed with human erythrocytes. Such biological activity was attributed to a certain proteic component that was purified by ion-exchange and size-exclusion chromatographies. The protein, so called camptosemin, has shown to occur as a tetramer and which oligomerizations state could be driven by the environmental pH. Primary sequence information from the amino-terminal portion, obtained by aminoacid sequencing, allowed the design of degenerated oligonucleotides for the amplification and cloning of its cDNA. The obtained clones sequences were submitted to several NCBI data bank search scripts. Among the retrieved results were found concanavalin and peanut agglutinin, two representatives of the called legume lectins family. Members of this group share with camptosemin their monomer sizes and the way their oligomerization state can be driven. This present work characterizes camptosemin with respect to its sequence and oligomerization using cloning and spectroscopy techniques. It was possible the cloning of two cDNA variants for camptosemin which sequences deduce an N-terminal signal peptide absent in the mature protein. Deduced primary sequences are very similar to each other and to other legume lectin members. Camptosemin has shown being extremely temperature resistant, only dissociating in monomeric subunits under extremely acid or alkaline pHs.
de, Oliveira Jose T. A. "Seed lectins : the effects of dietary Phaseolus vulgaris lectins on the general metabolism of monogastric animals". Thesis, University of Aberdeen, 1986. http://digitool.abdn.ac.uk/R?func=search-advanced-go&find_code1=WSN&request1=AAIU367276.
Pełny tekst źródłaOdom-Crespo, Eric William. "F-type lectins biochemical, genetic, and topological characterization of a novel lectin family in lower vertebrates /". College Park, Md. : University of Maryland, 2004. http://hdl.handle.net/1903/1437.
Pełny tekst źródłaThesis research directed by: Marine-Estuarine-Environmental Sciences. Title from t.p. of PDF. Includes bibliographical references. Published by UMI Dissertation Services, Ann Arbor, Mich. Also available in paper.
Lu, Jinhua. "Mammalian lectins containing collagen-like domains". Thesis, University of Oxford, 1991. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.303657.
Pełny tekst źródłaGupta, G. "Artificial lectins : biomimetic ligands for glycoproteins". Thesis, University of Cambridge, 2002. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.599789.
Pełny tekst źródłaSampaio, Alexandre Holanda. "Lectins from Ulva and Ptilota species". Thesis, University of Portsmouth, 1997. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.310389.
Pełny tekst źródłaSILVA, Flávio de Oliveira. "Atividade moduladora da lectina isolada das sementes de Canavalia brasiliensis". Universidade Federal Rural de Pernambuco, 2012. http://www.tede2.ufrpe.br:8080/tede2/handle/tede2/4585.
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Lectins are proteins that bind specifically and reversibly to carbohydrates, showing several biological effects. In this work, we carried out a literature review about biological effects of lectin extracted from seeds of Canavalia brasiliensis (ConBr), a plant present in the northeastern and southern Brazil, which is popularly known as wild bean of Ceará. In addition, we carried out a study to analyze the modulating activity of ConBr on murine splenocytes, verifying its effect on cell viability and proliferation, cytokine and nitric oxide (NO) production. We have also performed a study to evaluate ConBr effect on B16F10 murine melanoma cells by analyzing the inhibition of cell proliferation and migration as well as apoptosis induction and synthesis of cytokines and NO. The results show that ConBr induced at concentrations of 2.5, 5.0 and 10 μg/ml promoted the proliferation of splenocytes, with high cell viability. Furthermore, the concentration of 10 μg/ml induced cytokine production and nitric oxide on B16F10 murine melanoma, it was observed that ConBr inhibited tumor cell proliferation inducing apoptosis. It was also observed nitric oxide and IL-12 production by B16F10 cells under stimulus. ConBr lectin possesses a biotechnological potential use as a mitogen and anti-tumor agent.
As lectinas são proteínas que apresentam a capacidade de se ligar de maneira específica e reversível a carboidratos, exibindo distintos efeitos biológicos. Neste trabalho, realizou-se uma revisão de literatura sobre os efeitos biológicos da lectina extraída das sementes da Canavalia brasiliensis (ConBr), uma planta presente no Nordeste e Sul do Brasil, que é conhecida popularmente como feijão bravo do Ceará. Além disso, realizou-se um estudo para analisar a atividade moduladora da ConBr sobre esplenócitos murinos, verificando-se sua ação sobre a proliferação e viabilidade celular, produção de citocinas e óxido nítrico (NO). Realizou-se também, um estudo para avaliar o efeito da ConBr sobre células B16F10 de melanoma murino, analisando-se a inibição da proliferação e migração celular, bem como a indução de apoptose e síntese de citocinas e NO. Os resultados demostraram que a ConBr induziu nas concentrações de 2.5, 5.0 e 10 μg/ml promoveu a proliferação de esplenócitos, com alto índice de viabilidade celular. Além disso, a concentração de 10 μg/ml induziu a produção de citocinas e óxido nítrico. Em células B16F10 de melanoma murino, observou-se que a ConBr inibiu a proliferação das células tumorais promovendo apoptose celular. Verificou-se ainda, a produção de óxido nítrico e da citocina IL-12 pelas células submetidas ao estímulo. A lectina ConBr possue um potencial uso biotecnológico como mitógeno e agente antitumoral.
JÃnior, Francisco Nascimento Pereira. "PurificaÃÃo, caracterizaÃÃo parcial e cristalizaÃÃo de uma lectina ligante de manose das sementes de Platymiscium floribundum Vogel". Universidade Federal do CearÃ, 2011. http://www.teses.ufc.br/tde_busca/arquivo.php?codArquivo=5882.
Pełny tekst źródłaAs sementes de Platymiscium floribundum Vogel, uma espÃcie pertencente à famÃlia Leguminosae, subfamÃlia Papilionoideae, tribo Dalbergieae, possuem uma lectina manose/N-acetil-D-glicosamina especÃfica, que aglutina eritrÃcitos nativos ou tratados com enzimas proteolÃticas de coelho. A lectina de sementes de P. floribundum foi purificada por precipitaÃÃo com sulfato de amÃnio seguida por cromatografia de afinidade em matriz de Sepharose-manose. Esse procedimento resultou na lectina purificada, nomeada de PFL. O processo de purificaÃÃo da PFL foi monitorado por SDS-PAGE e atividade hemaglutinante especÃfica, observou-se que a lectina purificada à caracterizada por um perfil eletroforÃtico composto por uma Ãnica banda, com massa molecular aparente de aproximadamente 29 kDa, tanto na presenÃa quanto na ausÃncia de um agente redutor. A anÃlise por espectrometria de massa indicou que PFL possui massa molecular de 27.054  2 Da, e teve sua estrutura primaria parcialmente sequenciada atravÃs de espectrometria de massa sequencial. PFL à uma glicoproteÃna e demonstra elevada estabilidade, sendo capaz de manter sua atividade hemaglutinante apÃs exposiÃÃo a temperaturas de atà 60  C por 1 hora e na faixa de pH de 7,0 a 9,0. A PFL nÃo apresentou atividade anti-inflamatÃria.em modelo de edema de pata. PFL foi cristalizada em diferentes condiÃÃes de cristalizaÃÃo.
Platymiscium floribundum Vogel seeds, a species of the Leguminosae family, Papilionoideae subfamily, Dalbergieae tribe, have a lectin mannose/N-acetyl-D-glucosamine specific rabbit erythrocytes that agglutinate native or treated with proteolytic enzymes. The lectin from P. floribundum was purified by precipitation with ammonium sulfate followed by affinity chromatography on Sepharose-mannose. This procedure resulted in a purified lectin, named PFL. PFL purification process was monitored by SDS-PAGE and showed that the purified lectin is characterized by an electrophoretic profile consists of a single band with apparent molecular mass of approximately 29 kDa, in both presence and absence of an reducer agent. The analysis by mass spectrometry indicated that PFL has a molecular mass of 27,054 Da, and its primary structure was partially sequenced. PFL is a glycoprotein and shows high stability, being able to maintain its haemagglutinating activity after exposure to temperatures up to 60 ÂC for one hour and at pH 7.0 to 9.0. The PFL showed no anti-inflammatory activity in paw edema model. PFL was crystallized under different crystallization.
Landim, PatrÃcia Gadelha de Castro. "Xiloglucanas e galactomananas de leguminosas: interaÃÃo com lectinas D-galactose-ligantes". Universidade Federal do CearÃ, 2007. http://www.teses.ufc.br/tde_busca/arquivo.php?codArquivo=8583.
Pełny tekst źródłaPolissacarÃdeos ocorrem em grandes quantidades nas sementes como componentes da parede celular ou como reserva. Dentre estes Ãltimos, incluem-se as xiloglucanas cotiledonÃrias e as galactomananas endospÃrmicas. As xiloglucanas apresentam uma cadeia principal de β-D-(1→4)-glucana ramificada com ligaÃÃes α-(1→6) por resÃduos D-xilopiranosÃdeos ou β-D-galactopiranosÃdeo-(1→2)-D-xilopiranosÃdeos, enquanto as galactomananas endospÃrmicas consistem em cadeias polimÃricas de resÃduos β-D-manopiranosil (1→4) ligados, substituÃdos em O-6 por unidades de α-D-galactopiranosil. O objetivo deste trabalho foi analisar a interaÃÃo de xiloglucanas e galactomananas com lectinas galactose-ligantes e, assim, sugerir o uso de tais polissacarÃdeos como alternativa barata e eficaz na preparaÃÃo de matrizes cromatogrÃficas para o isolamento e determinaÃÃo da especificidade anomÃrica de novas lectinas. Dessa forma, as interaÃÃes das lectinas das sementes de Artocarpus integrifolia (frutalina), Artocarpus incisa (jacalina), Ricinus communis (ricina) e Arachis hypogaea (PNA) com matrizes de xiloglucanas de sementes de Copaifera langsdorffii, Mucuna sloanei e Hymenaea courbaril (MC, MMu, MJ, respectivamente) e de galactomananas de Mimosa scabrella, Stryphnodendron barbatiman, Adenanthera pavonina e Dimorphandra mollis (MM, MS, MA; MD, respectivamente) foram analisadas. As galactomananas apresentaram melhor capacidade de retenÃÃo da jacalina (MA â 0,92 mg ; MM â 1,48 mg ; MD â0,88 mg ; MS â 0,83 mg) e da frutalina (MA â 0,99 mg ; MM â 1,09 mg ; MD â 0,94 mg ; MS â 0,85 mg), lectinas que possuem especificidade por α-D-galactose. Vale destacar que a galactomanana de M. scabrella apresentou melhor capacidade de retenÃÃo da lectinas testadas. Por outro lado a ricina, capaz de ligar-se aos dois anÃmeros, mas que se liga preferencialmente ao anÃmero β, teve maior massa retida nas colunas de xiloglucana (MMu â 2,17 mg ;MJ â 1,30 mg; MC â 2,83 mg). Houve diferenÃa nos perfis de retenÃÃo da PNA, que tambÃm se liga aos anÃmeros α e β da galactose, sendo que a melhor retenÃÃo foi na coluna contendo matriz de M. sloanei (0,12 mg). As colunas foram todas saturadas com extrato bruto a partir das farinhas das sementes, para que se utilizasse a capacidade mÃxima de retenÃÃo de cada matriz. Atividade hemaglutinante foi detectada em ambos os picos PI e PII. Para a ricina, atividade tÃxica foi realizada e detectada para todos os PII obtidos. Por meio de SDS-PAGE, a pureza de cada uma das lectinas foi confirmada. Diante dos resultados expostos, pode-se sugerir o uso de xiloglucanas e galactomananas para o isolamento, purificaÃÃo e determinaÃÃo da especificidade anomÃrica de lectinas galactose-ligantes.
Polysaccharides are found in large quantity in seeds and they represent the main compounds of cell wall or reservoir. Among reservoir compounds, it included cotyledonary xyloglucans and endospermic galactomannans. The xyloglucans are made of a main chain of β-D-(1→4)-glucan with α-(1→6) ramifications of D-xylopyranoside or β-D-galactopyranoside-(1→2)-D-xylopyranoside residues. Endospermic galactomannans are polimeric chains of β-D-mannopyranosil (1→4) and replaceabled in O-6 for units of α-D-galactopyranosil. The aim of this work is investigate the interaction of xyloglucans and galactomannans with galactose bounding lectins and show the possibility of the usage of these polysaccharides as cheap and useful chromatographic matrices for isolation and determination of anomeric specificity of galactose bounding lectins. The interactions of lectins from seeds of Artocarpus integrifolia (frutalin), Artocarpus incisa (jacalin), Ricinus communis (ricin) e Arachis hypogaea (PNA) were performed with coluns of xyloglucans of seeds from Copaifera langsdorffii, Mucuna sloanei and Hymenaea courbaril (MC, MMu, MJ, respectively) and galactomannans from Mimosa scabrella, Stryphnodendron barbatiman, Adenanthera pavonina and Dimorphandra mollis (MM, MS, MA; MD, respectively). The galactomannans showed the best colun interaction capacity for the jacalin (MA â 0,92 mg ; MM â 1,48 mg ; MD â0,88 mg ; MS â 0,83 mg) and frutalin (MA â 0,99 mg ; MM â 1,09 mg ; MD - 0,94 mg ; MS - 0,85 mg) lectins. Remarkably the M. scabrella galactomannan showed the best colun interaction among all lectins analysed. On the other hand, ricin was better hold in coluns made of xyloglucan (MMu â 2,17 mg ;MJ â 1,30 mg; MC â 2,83 mg). For PNA lectin, differences were detected in colun interaction capacity. The best colun interaction was with the M. sloanei matrix (0,12 mg) for PNA lectin. All coluns were fill with sample extract of flour from seeds and hemagglutination assays was performed with PI and PII. In these assays, hemagglutination activity was detected in both PI and PII from the coluns. For ricin, toxic activity was made and it was detected for all obtained chromatographic samples. With SDS-PAGE it was possible confirmed the purification of the studied lectins. The bands in polyacrilamid gel were the same for the lectins purified. In conclusion, it can be suggested the usage of xyloglucans and galactomannans for isolation, purification and determination of anomeric specificity of galactose-bounding lectins.
Lobo, Marina Duarte Pinto. "AnÃlise proteÃmica de plasma de pacientes com cÃncer de mama utilizando lectinas vegetais e label-free LC-MSE". Universidade Federal do CearÃ, 2016. http://www.teses.ufc.br/tde_busca/arquivo.php?codArquivo=16410.
Pełny tekst źródłaO cÃncer de mama representa o tipo de cÃncer mais comum entre as mulheres no mundo e no Brasil, depois do cÃncer de pele nÃo melanoma. Caracterizando-se como uma doenÃa clÃnica e biologicamente heterogÃnea, a detecÃÃo precoce do cÃncer de mama, uma caracterizaÃÃo fidedigna da doenÃa e um diagnÃstico preciso sÃo de fundamental importÃncia para reduÃÃo da mortalidade. Assim, o objetivo do presente trabalho foi realizar anÃlises qualitativas e quantitativas de proteÃnas plasmÃticas de mulheres saudÃveis e mulheres com cÃncer de mama tipo ductal, em diferentes estÃgios. Para isso, inicialmente, as amostras de plasma foram depletadas de albumina e IgG e depois reunidas em pools representativos para cada grupo em estudo. Os pools foram entÃo fracionados por meio de cromatografias de afinidade em matrizes de Sepharose 4B imobilizadas com as lectinas vegetais α-galactose-ligante de Artocarpus incisa - Frutalina e glucose/manose-ligante de Dioclea altÃssima â DAL. Posteriormente, tantos os pools como as fraÃÃes cromatogrÃficas obtidas foram digeridas e submetidas à anÃlise independente de dados (MSE) e quantificadas (label-free) por espectrometria de massas. A utilizaÃÃo das cromatografias de afinidade com lectinas vegetais a priori da anÃlise por espectrometria de massas foi fundamental para reduzir a complexidade das amostras e estender o intervalo dinÃmico de proteÃnas, e frutalina apresentou os melhores resultados de fracionamento. Um somatÃrio de todas as anÃlises realizadas revelou a identificaÃÃo de cerca de 445.000 peptÃdeos e mais de 30.000 proteÃnas, com redundÃncia entre isoformas do mesmo grupo e entre grupos. Ademais, alÃm de fracionar, as cromatografias de afinidade com lectinas vegetais foram eficientes em isolar glicoformas especÃficas que refletem um padrÃo de glicosilaÃÃo alterado associado à doenÃa. Diversas proteÃnas diferencialmente expressas, algumas delas com mudanÃas na glicosilaÃÃo, foram identificadas, tais como apolipoproteÃna A2, apolipoproteÃna C3, fatores do sistema complemento (C3, C4b e C4A), clusterina, α-1-2-Ãcido glicoproteÃna, haptoglobina, hemopexina, paraoxonase arilesterase sÃrica, proteÃna plasmÃtica de ligaÃÃo ao retinol, plasminogÃnio e vitronectina. Dentre as proteÃnas identificadas, algumas estÃo relacionadas com a decomposiÃÃo da matriz extracelular, metabolismo lipÃdico, estresse oxidativo e outras caracterizam-se como proteÃnas de fase aguda. Finalmente, os dados de expressÃo diferencial e possÃveis alteraÃÃes de glicosilaÃÃo das proteÃnas contribuem para o desenvolvimento de um perfil protÃico, alvo para posterior validaÃÃo, que apresenta uma associaÃÃo com o desenvolvimento e a caracterizaÃÃo do cÃncer de mama em seus diferentes estÃgios.
The breast cancer presents one of the most commonly diagnosed types of cancer among women worldwide and in Brazil, after nonmelanoma skin cancer. Itâs a clinically and biologically heterogeneous disease. Therefore, early detection of breast cancer, a reliable characterization of the disease and an accurate diagnosis are crucial to reducing mortality. The aim of this work was perform a qualitative and quantitative analyzes of plasma proteins from healthy women and from women with ductal breast cancer in different stages. For this, initially, plasma samples were depleted of IgG and albumin and then pooled into samples representative for each study group. The pools were then fractionated by immobilized plant lectins (frutalin and DAL)-affinity chromatography. Subsequently, the pools and chromatographic fractions were digested and submited to and data-independent label-free mass spectrometric analysis. The use of affinity chromatography with plant lectins prior to analysis by mass spectrometry was essential to reduce sample complexity and extend the dynamic range of proteins, and frutalin showed the best results from fractionation. A sum of all analyzes performed revealed the identification of about 445,000 peptides and more than 30,000 proteins, with redundancy between isoforms of the same group and between groups. Furthermore, in addition to fractionation, the chromatography of affinity with plant lectins were effective in isolating specific glycoforms that reflect an altered glycosylation pattern associated with the disease. Several differentially expressed proteins, some of which changes in glycosylation were identified, such as apolipoprotein A2, apolipoprotein C3, complement factors (C3, C4b and C4A), clusterin, 1-2-α-acid glycoprotein, haptoglobin, hemopexin, serum paraoxonase arylesterase, plasma retinol binding protein, plasminogen and vitronectin. Among the proteins identified, some are related to the breakdown of the extracellular matrix, lipid metabolism, oxidative stress and other characterized as acute phase proteins. Finally, the data of differential expression and possible changes in the glycosylation patterns of proteins contributes to the development of a protein profile, subject to later validation, presenting an association with the development and characterization of breast cancer in its different stages.
Sousa, Bruno Lopes de. "Caracterização estrutural das formas silvestre e recombinante de uma lectina de sementes de Vatairea macrocarpa Benth e análise das suas bases moleculares de ligação ao antígeno Tn". reponame:Repositório Institucional da UFC, 2014. http://www.repositorio.ufc.br/handle/riufc/18875.
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Lectins are a very diverse class of proteins able to bind specific sugar structures reversibly and with high specificity, but without enzymatically modifying them, triggering several important cellular processes. Among the different lectin families, legume lectins are the most thoroughly studied and have been widely reported to exhibit a number of links to many pathological processes, including carcinogenesis. The remarkable anti-tumor properties of some legume lectins, resulting from their ability to induce programmed cell death and/or autophagocytosis in cancer cells have attracted much attention for biomedical applications. Moreover, a few galactose/N-acetylgalactosamine (Gal/GalNAc)-binding lectins from this group have proven to be useful markers for cancer histochemistry, and the structural characterization of these lectins bound to specific cancer epitopes has been carried out successfully. The seed lectin isolated from the legume tree Vatairea macrocarpa (VML) is a well characterized Gal/GalNAc-binding protein able to specifically recognize naturally occurring O-mucins presenting the carcinoma epitope Tn antigen (GalNAc-α-O-Ser). The crystal structures of VML in complex with Tn antigen and GalNAc have been determined at the resolution of 1.4 and 1.7 Å, respectively. Unfortunately, most of lectins obtained from natural sources consist in a mixture of forms, which is an undesired feature for biomedical applications. Thus, the recombinant form of VML (rVML) was expressed in Escherichia coli, being obtained soluble and wih high yielding. The crystal structure for rVML, as well as for the complex with Tn antigen, GalNAc and α-Lactose have been determined at resolutions of 1.7, 2.7, 2 and 1.8 Å, respectively, presenting the same overall structure and binding patterns as the wild lectin. Molecular docking analysis of this new structure and other Tn-binding legume lectins to O-mucin fragments differently decorated with this antigen provides a comparative binding profile among these proteins, stressing that subtle alterations that may not influence monosaccharide binding can, nonetheless, directly impact the ability of these lectins to recognize naturally occurring antigens. In addition to the specific biological effects of VML, the structural and binding similarities between it and other lectins commonly used as histological markers (e.g., VVLB4 and SBA) strongly suggest that VML can be used as a new tool for cancer research. This is the first report of crystal structures of a Gal/GalNAc-binding legume lectin from the Dalbergieae tribe.
As lectinas consitem em uma classe diversificada de proteínas, capazes de reconhecer estruturas glicídicas de forma reversível e com alta especificidade, no entanto sem alterar suas estruturas químicas, participando de vários processos celulares importantes. Dentre as diferentes famílias de lectinas, as isoladas a partir de leguminosas são as mais extensivamente estudadas, havendo sido relatada a influências dessas moléculas sobre diversos processos patológicos, incluindo a carcinogênese. Notáveis propriedades antitumorais têm sido detectadas para algumas lectinas de leguminosas, resultantes da sua habilidade em induzir a morte celular ou a autofagia em células cancerígenas, o que atraído atenção para suas possiveis aplicações biomédicas. Além disso, algumas lectinas desse grupo especificas para galactose/N-acetil-D-galactosamina (Gal/GalNAc) têm se mostrado úteis como marcadores histoquímicos na pesquisa do câncer e a caracterização estrutural dessas lectinas em complexo com diferentes epítopos cancerígenos vem sendo realizada com sucesso. A lectina isolada a partir das sementes da leguminosa Vatairea macrocarpa (VML) é uma lectina bem caracterizada específica para Gal/GalNAc capaz de reconhecer especificamente o antígeno Tn (GalNAc-α-O-Ser), naturalmente encontrado em O-mucinas presentes em diferentes tipos de câncer. As estruturas cristalográficas para a VML em complexo com o antígeno Tn e GalNAc foram determinadas com resoluções de 1.4 e 1.7 Å, respectivamente. A maioria das lectinas obtidas a partir de fontes naturais consiste em misturas de diferentes isoformas, uma característica indesejada para aplicações biomédicas. Com base nisso, uma construção recombinante para VML (rVML) foi expressa em Escherichia coli, sendo obtida de forma solúvel em com alto rendimento. A estrutura cristalina para a rVML, bem como para seus complexos com o antígeno Tn, GalNAc e α-Lactose foram determinadas com resoluções de 1.7, 2.7, 2 e 1.8 Å, respectivamente, apresentando a mesma estrutura geral e padrões de interação que a lectina silvestre. Com o intuito de gerar um perfil comparativo entre a VML e outras lectinas de leguminosas capazes de reconhecer o antígeno Tn, foram realizadas análises de docking molecular utilizando fragmentos de O-mucinas diferentemente decorados com o antígeno Tn. Esse perfil ressalta como alterações sutis no elenco ou disposição dos aminoácidos constituintes do sítio de ligação a carboidrato, que talvez não influenciem a capacidade de ligação a monossacarídeo, podem impactar diretamente a habilidade dessas lectinas em reconhecer antígenos em condições naturais. Adicionalmente aos já caracterizados efeitos biológicos relatados para VML, a similaridade entre sua estrutura e perfis de interações quando comparadas a outras lectinas comumente utilizadas como marcadores histoquímicos (e.g., VVLB4 e SBA), sugerem fortemente a possível utilização da VML como uma nova ferramenta na pesquisa do câncer. Esse trabalho consiste no primeiro relato de estruturas cristalográficas para uma lectina de leguminosa específica para Gal/GalNAc da tribo Dalbergieae.
Landim, Patrícia Gadelha de Castro. "Xiloglucanas e galactomananas de leguminosas: interação com lectinas D-galactose-ligantes". reponame:Repositório Institucional da UFC, 2007. http://www.repositorio.ufc.br/handle/riufc/18846.
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Polysaccharides are found in large quantity in seeds and they represent the main compounds of cell wall or reservoir. Among reservoir compounds, it included cotyledonary xyloglucans and endospermic galactomannans. The xyloglucans are made of a main chain of β-D-(1→4)-glucan with α-(1→6) ramifications of D-xylopyranoside or β-D-galactopyranoside-(1→2)-D-xylopyranoside residues. Endospermic galactomannans are polimeric chains of β-D-mannopyranosil (1→4) and replaceabled in O-6 for units of α-D-galactopyranosil. The aim of this work is investigate the interaction of xyloglucans and galactomannans with galactose bounding lectins and show the possibility of the usage of these polysaccharides as cheap and useful chromatographic matrices for isolation and determination of anomeric specificity of galactose bounding lectins. The interactions of lectins from seeds of Artocarpus integrifolia (frutalin), Artocarpus incisa (jacalin), Ricinus communis (ricin) e Arachis hypogaea (PNA) were performed with coluns of xyloglucans of seeds from Copaifera langsdorffii, Mucuna sloanei and Hymenaea courbaril (MC, MMu, MJ, respectively) and galactomannans from Mimosa scabrella, Stryphnodendron barbatiman, Adenanthera pavonina and Dimorphandra mollis (MM, MS, MA; MD, respectively). The galactomannans showed the best colun interaction capacity for the jacalin (MA – 0,92 mg ; MM – 1,48 mg ; MD –0,88 mg ; MS – 0,83 mg) and frutalin (MA – 0,99 mg ; MM – 1,09 mg ; MD - 0,94 mg ; MS - 0,85 mg) lectins. Remarkably the M. scabrella galactomannan showed the best colun interaction among all lectins analysed. On the other hand, ricin was better hold in coluns made of xyloglucan (MMu – 2,17 mg ;MJ – 1,30 mg; MC – 2,83 mg). For PNA lectin, differences were detected in colun interaction capacity. The best colun interaction was with the M. sloanei matrix (0,12 mg) for PNA lectin. All coluns were fill with sample extract of flour from seeds and hemagglutination assays was performed with PI and PII. In these assays, hemagglutination activity was detected in both PI and PII from the coluns. For ricin, toxic activity was made and it was detected for all obtained chromatographic samples. With SDS-PAGE it was possible confirmed the purification of the studied lectins. The bands in polyacrilamid gel were the same for the lectins purified. In conclusion, it can be suggested the usage of xyloglucans and galactomannans for isolation, purification and determination of anomeric specificity of galactose-bounding lectins.
Polissacarídeos ocorrem em grandes quantidades nas sementes como componentes da parede celular ou como reserva. Dentre estes últimos, incluem-se as xiloglucanas cotiledonárias e as galactomananas endospérmicas. As xiloglucanas apresentam uma cadeia principal de β-D-(1→4)-glucana ramificada com ligações α-(1→6) por resíduos D-xilopiranosídeos ou β-D-galactopiranosídeo-(1→2)-D-xilopiranosídeos, enquanto as galactomananas endospérmicas consistem em cadeias poliméricas de resíduos β-D-manopiranosil (1→4) ligados, substituídos em O-6 por unidades de α-D-galactopiranosil. O objetivo deste trabalho foi analisar a interação de xiloglucanas e galactomananas com lectinas galactose-ligantes e, assim, sugerir o uso de tais polissacarídeos como alternativa barata e eficaz na preparação de matrizes cromatográficas para o isolamento e determinação da especificidade anomérica de novas lectinas. Dessa forma, as interações das lectinas das sementes de Artocarpus integrifolia (frutalina), Artocarpus incisa (jacalina), Ricinus communis (ricina) e Arachis hypogaea (PNA) com matrizes de xiloglucanas de sementes de Copaifera langsdorffii, Mucuna sloanei e Hymenaea courbaril (MC, MMu, MJ, respectivamente) e de galactomananas de Mimosa scabrella, Stryphnodendron barbatiman, Adenanthera pavonina e Dimorphandra mollis (MM, MS, MA; MD, respectivamente) foram analisadas. As galactomananas apresentaram melhor capacidade de retenção da jacalina (MA – 0,92 mg ; MM – 1,48 mg ; MD –0,88 mg ; MS – 0,83 mg) e da frutalina (MA – 0,99 mg ; MM – 1,09 mg ; MD – 0,94 mg ; MS – 0,85 mg), lectinas que possuem especificidade por α-D-galactose. Vale destacar que a galactomanana de M. scabrella apresentou melhor capacidade de retenção da lectinas testadas. Por outro lado a ricina, capaz de ligar-se aos dois anômeros, mas que se liga preferencialmente ao anômero β, teve maior massa retida nas colunas de xiloglucana (MMu – 2,17 mg ;MJ – 1,30 mg; MC – 2,83 mg). Houve diferença nos perfis de retenção da PNA, que também se liga aos anômeros α e β da galactose, sendo que a melhor retenção foi na coluna contendo matriz de M. sloanei (0,12 mg). As colunas foram todas saturadas com extrato bruto a partir das farinhas das sementes, para que se utilizasse a capacidade máxima de retenção de cada matriz. Atividade hemaglutinante foi detectada em ambos os picos PI e PII. Para a ricina, atividade tóxica foi realizada e detectada para todos os PII obtidos. Por meio de SDS-PAGE, a pureza de cada uma das lectinas foi confirmada. Diante dos resultados expostos, pode-se sugerir o uso de xiloglucanas e galactomananas para o isolamento, purificação e determinação da especificidade anomérica de lectinas galactose-ligantes.
Kohatsu, Luciana. "Novel roles for human lectins in innate defense against pathogens". Diss., Restricted to subscribing institutions, 2006. http://proquest.umi.com/pqdweb?did=1155572661&sid=1&Fmt=2&clientId=1564&RQT=309&VName=PQD.
Pełny tekst źródłaJohansson, Emma M. V. "Combinatorial discovery of glycopeptide dendrimers targeting lectins /". [S.l.] : [s.n.], 2009. http://www.ub.unibe.ch/content/bibliotheken_sammlungen/sondersammlungen/dissen_bestellformular/index_ger.html.
Pełny tekst źródłaVieira, Breitwieser Otilia. "Leguminous lectins bind non specifically to DNA". [S.l. : s.n.], 2003. http://deposit.ddb.de/cgi-bin/dokserv?idn=972705589.
Pełny tekst źródłaHutchinson, Oliver Clyde. "The isolation and characterization of fungal lectins". Thesis, Queen's University Belfast, 2000. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.322949.
Pełny tekst źródłaAl-Abbasi, Fahad Ahmed. "Application of lectins to specific analytical problems". Thesis, University of Newcastle Upon Tyne, 2007. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.438401.
Pełny tekst źródłaCarter, Tom Scott. "Synthetic lectins with pyrene at the core". Thesis, University of Bristol, 2016. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.702443.
Pełny tekst źródłaBarroso, Neto Ito Liberato. "Resolução da estrutura tridimensional de uma lectina de sementes de Canavalia grandiflora Benth. com efeitos sobre mecanismos inflamatórios". reponame:Repositório Institucional da UFC, 2010. http://www.repositorio.ufc.br/handle/riufc/18171.
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Plant Lectins can be defined as non-immune origin proteins having at least one non-catalytic domain that binds reversibly in a specific fashion to a mono-or oligosaccharide. The legume lectin family group represents the best studied of these proteins, in particular highlighted the subtribe Diocleinae, which has the best characterized representative, ConA. Lectins Diocleinae show a high degree of structural similarity, but the same does not regarding biological activities and specificity to different carbohydrates, which makes it important to research in structural and biological levels of its various members, among them the Canavalia grandiflora Benth. The lectin from Canavalia grandiflora (Congr) was purified according to Ceccato (2001) and was crystallized by vapor diffusion method at 293 K. Crystals were obtained in a state containing 0.5 M of cadmium sulfate hydrate, 0.1 M HEPES pH 7.5 and 1.0 M sodium acetate trihydrate. The crystals have the orthorhombic space group I222, the unit cell has the dimensions a = 67.70 Å, b = 55.90 Å and c = 107.46 Å and angles? =? =? = 90 °, giving a monomer in the asymmetric unit and a content of 42.53% solvent in the crystal. The structure was solved to 2.19 Å and phase problem was solved by the method of molecular replacement using the coordinates of Canavalia gladiata as a template (PDB: 2D7F). The refinement showed satisfactory "rfactor" and "Rfree" with 22.6 and 27.4 respectively and only one amino acid residue in a region of the Ramachandran disallowed. Despite the high structural similarity, small changes in the direction of key amino acids may be responsible for the diversity in the biological applications as a result of the residues of the carbohydrate binding site which are retained despite changes in spatial orientation. The primary sequence of the lectin from C. grandiflora has great similarity with lectins of the same genus, but it has the largest number of mutations representative of the genus Dioclea, characterizing it as the subgenus Canavalia nearest Dioclea and canavalias is among the most primitive. The Congregation edamatogênica showed activity in a model of paw edema (sc) and relaxing effect on smooth muscle of rat aorta endothelial, but the effects appear to be weak compared to other lectins Diocleinae.
Lectinas de plantas podem ser definidas como proteínas de origem não imune que possuem pelo menos um domínio não catalítico que se liga reversivelmente de maneira específica a um mono- ou oligossacarídeo. A família das lectinas de leguminosas representa o grupo destas proteínas mais bem estudadas, em especial destaque a subtribo Diocleinae, que possui o representante mais bem caracterizado, a ConA. As lectinas de Diocleinae apresentam um alto grau de similaridade estrutural, porém o mesmo não ocorre quanto às atividades biológicas e especificidade a carboidratos variados, o que torna importante a investigação em níveis estruturais e biológicos dos seus diversos membros, dentre eles a Canavalia grandiflora Benth. A lectina de sementes de Canavalia grandiflora (ConGr) foi purificada de acordo com Ceccato (2001) e foi cristalizada pelo método de difusão de vapor a 293 K. Cristais foram obtidos em uma condição contendo 0,5 M de sulfato de cádmio hidratado, 0,1 M de HEPES pH 7.5 e 1,0 M de acetato de sódio triidratado. Os cristais apresentam o grupo espacial ortorrômbico I222, a cela unitária tem como dimensões a=67,70 Å, b=55,90Å e c=107,46 Å e ângulos ?=?=?=90º, sendo observado um monômero na unidade assimétrica e um conteúdo de 42,53% de solvente no cristal. A estrutura foi resolvida a 2,19Å e o problema de fase foi solucionado pelo método de substituição molecular utilizando as coordenadas da Canavalia gladiata como modelo (PDB: 2D7F). O refinamento satisfatório apresentou “Rfactor” e “Rfree” com 22,6 e 27,4 respectivamente e apenas um resíduo de aminoácido em região não permitida do Ramachandran. Apesar da alta similaridade estrutural, pequenas mudanças na orientação de aminoácidos chaves, podem ser responsáveis pela diversidade nos resultado de aplicações biológicas como os resíduos do sítio de ligação a carboidrato, que apesar de conservados possuem modificações na orientação espacial. A seqüência primária da lectina de C. grandiflora apresenta grande similaridade com lectinas do mesmo gênero, porém ela concentra o maior número de mutações representativas do gênero Dioclea, caracterizando-o como o subgênero de Canavalia mais próximo de Dioclea, e dentre as canavalias é a mais primitiva. A ConGr apresentou atividade edamatogênica em modelo de edema de pata (s.c.) e efeito relaxante em músculo liso de aortas de ratos endotelizadas, porém os efeitos mostram-se fracos frente a outras lectinas de Diocleinae.
Oliveira, Mara Rubea Tinoco Rodrigues de. "Avaliação in vitro do efeito de lectinas de sementes de Talisia esculenta e Labramia bojeri sobre o biofilme oral". [s.n.], 2005. http://repositorio.unicamp.br/jspui/handle/REPOSIP/288965.
Pełny tekst źródłaDissertação (mestrado profissional) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba
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Resumo: A medicina natural e complementar, especialmente a fitoterapia, suprem as necessidades em saúde de grande parte da população, particularmente nos países em desenvolvimento. Dentre os fitoterápicos, as lectinas podem ter valia como agentes antiplaca, uma vez que podem estar intimamente relacionadas com a aderência de microrganismos. O objetivo deste estudo foi testar in vitro a capacidade de inibição das lectinas isoladas (TEL - derivada da semente de Talisia esculenta e LABRAMIN - purificada da planta Labramia bojeri), na adesão e crescimento de microrganismos orais (Streptococcus sanguinis, S. mitis, S. oralis, S. mutans, S. sobrinus). A atividade antimicrobiana das duas lectinas foi determinada pelo teste convencional da macrodiluição de caldo, sendo testada as concentrações de 400, 200, 100, 50, 25 µg/mL contra 105 CFU/mL dos microrganismos em estudo. Os tubos foram incubados (10% CO2, 37oC, 18h) e submetidos à leitura de densidade óptica (OD a 600nm). A MBC foi determinada pela adição das amostras de cada tubo em placas de petri contendo agar BHI (10% CO2, 37oC, 18h). Para avaliação de aderência foi feito um ensaio semiquantitativo de aderência em placas de microtitulação de poliestireno, onde foi adicionado 100 µL de saliva clarificada e incubada por 2h a 37°. Após lavagem com PBS, foram adicionadas às placas em triplicata 100 µL de lectinas (6.25, 12.5, 25, 50 e 100 µg/mL) e incubados durante 1h. A seguir foi adicionada 100 µL de suspensão bacteriana (106 UFC/mL) e incubado a 37ºC em uma atmosfera de 10% de CO2. A aderência foi revelada e quantificada por tintura com cristal violeta. A absorção do cristal violeta foi determinada por um leitor de placa (575nm). Nem a TEL nem a LABRAMIN foram capazes de inibir ou matar os microorganismos estudados. No teste de inibição de aderência, a LABRAMIN reduziu significativamente a aderência de S. mutans e S. sobrinus, na concentração de 100µg (p< 0,05). A TEL não inibiu a aderência dos estreptococos e ainda favoreceu a aderência do S. mitis. Concluiu-se que embora nenhuma das lectinas estudadas tenham atividade antimicrobiana, a LABRAMIN foi capaz de inibir a aderência de estreptococos cariogênicos
Abstract: Natural and complementary medicine, with special regard to phytotherapy, provide care for health needs of a great part of the world¿s population, particularly in developing countries. Among phytotherapic compounds, lectins could be of value as anti-plaque agents, once they can be closely related to microorganisms¿ adherence. Our study intended to test, in vitro, the capacity of two isolated lectins ¿ TEL (derived from Talisia esculenta seeds) and LABOL (purified from the plant Labramia bojeri ¿ for inhibiting the adherence and growth of oral microorganisms (Streptococcus sanguinis, S. mitis, S. oralis, S. mutans, S. sobrinus). Both lectins¿ antimicrobial activities were determined by a conventional macrodilution broth test. Study¿s microorganisms concentrations were tested for 400, 200, 100, 50, 25 µg/mL against 105 CFU/mL. Tubes were incubated (10% CO2, 37oC, 18h) and submitted to optical density reading (OD at 600nm). MBC was determined by the addition of samples from each tube to petri dishes containing agar BHI (10% CO2, 37oC, 18h). A semiquantitative assay was performed for adherence assessment in polystyren microtiter plates. A hundred microliter (100µL) of clarified saliva were added and incubated for 2h at 37°. Following PBS wash, 100 µL lectins (6.25, 12.5, 25, 50 e 100 mg/mL) were added to the triple plates and incubated for 1h. Then, 100 µL of bacterial suspension (106 UFC/mL) were added and incubated at 37ºC in a 10% CO2 atmosphere. Adherence was revealed and quantified by crystal violet staining. A plate reader (575nm) determined crystal violet absorbance. Nor TEL nor LABOL were able to inhibit or kill the studied microorganisms. Regarding adherence inhibition, 100µg concentration of LABOL showed statistically significant (p < 0.05) on S.mutans and S. sobrinus. TEL did not inhibt streptococci adherence; yet, it favored S. mitis adherence. It was concluded that, although none of the studied lectins had presented antimicrobial activity, LABOL was capable of inhibiting cariogenic streptococci adherence. Further studies will be necessary to assess such lectins¿ potential over other microorganisms and over oral biofilm
Mestrado
Mestre em Odontologia em Saúde Coletiva
Silveira, Willian Abraham da. "Avaliação da interação entre galectina-1 e zinco e suas potenciais implicações estruturais e funcionais". Universidade de São Paulo, 2011. http://www.teses.usp.br/teses/disponiveis/60/60135/tde-05072011-122646/.
Pełny tekst źródłaIntroduction: Galectin-1 (Gal-1) is a multifunctional protein that specifically recognizes glycans with -galactosides through carbohydrate recognition domains (CRD). Gal-1 is a homodimeric protein of 14.900daltons, pI=5.6, shows a jelly-roll molecular topology composed of two anti-parallels - sheet, has no signal peptide and contains 6 cysteines, 7 glutamic acids, 9 aspartic acids and 4 histidines per monomer. This lectin binds to different biological molecules contained in the cell surface, nucleus and extracellular matrix components. Zinc is an important metal in biological systems because can participate in the maintenance of protein structure and biological activity. Usually, cysteine , histidine, glutamic acid and aspartic acid residues are preferential targets for interaction with Zn. Approximately 10% of the human proteome is potentially capable to forming complexes with Zn. The Zn2+ ion exhibits properties suitable for both catalytic and structural protein functions. Proteins zinc binding sites can be divided into catalytic, structural, co-catalytic and protein interface sites.There are reports in the literature that shows the interaction between galectin-1 and organic ions. However, were not found reports about Zn-Gal-1 complexes. Objective: The aim of this study was to evaluate the existence and implications of the interaction between galectin-1 and Zn2+ ion. Materials and Methods: Human recombinant Gal-1 (monomer and dimmer) was obtained and purified. Also, the conditions for the use of Gal-1 were standardized. The interaction Zn/Gal-1 was assessed by biophysical an biological procedures. The analysis in vitro and in silico was made by spectrofluorimetry, circular dichroism, precipitation test, method of GRID, and molecular dynamics. The in vitro analysis of biological parameters were performed by hemmaglutination and laminin binding (ELISA) tests. Results and Discussion: The addition of ZnCl2 in Gal-1 solution causes increased fluorescence emission of tryptophan-70 and a red shift, alters the circular dichroism spectrum and causes precipitation of Gal-1 protein. These events occurred in a selective manner dependent of Zinc concentration. The in silico analysis indicates that the probable site of Zn/Gal-1 complexation is distinct from the CRD and is formed by the amino acids Glu-15, Asp-92 and Asp-134, assuming trigonal bipyramidal conformation and with coordination number equal to 5 . Conclusion: The biophysical in vitro and in silico findings suggests that Galectin-1 has the ability to complex with the Zn2+ ion.
Carding, S. R. "Immunochemical studies of mammalian beta-galactoside ?-binding lectins". Thesis, University of Cambridge, 1985. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.352601.
Pełny tekst źródłaBarwell, Nicholas P. "Synthetic lectins : biomimetic receptors for carbohydrates in water". Thesis, University of Bristol, 2009. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.495920.
Pełny tekst źródłaRaemaekers, Romaan J. M. "Expression of functional plant lectins in heterologous systems". Thesis, Durham University, 2000. http://etheses.dur.ac.uk/4621/.
Pełny tekst źródłaWoodhouse, Stephen David. "Binding and toxicity of plant lectins to insects". Thesis, Durham University, 2002. http://etheses.dur.ac.uk/4103/.
Pełny tekst źródłaFontinopoulou, Angeliki. "Investigating the glycosylation of recombinant glycoproteins with lectins". Thesis, University of Newcastle Upon Tyne, 2003. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.432779.
Pełny tekst źródłaNantwi, Paul Kwasi Kankam. "Lectins in drug delivery to the oral cavity". Thesis, University of Portsmouth, 1997. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.482091.
Pełny tekst źródłaRenney, Charles M. "Novel synthetic lectins for carbohydrate recognition in water". Thesis, University of Bristol, 2015. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.686165.
Pełny tekst źródłaKaspar, Amir Shermila. "Synthetic lectins with ethylene spacers recognition in water". Thesis, University of Bristol, 2017. https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.738277.
Pełny tekst źródłaZelensky, Alex N. "In silico analysis of C-type lectin domains' structure and properties /". View thesis entry in Australian Digital Theses Program, 2004. http://thesis.anu.edu.au/public/adt-ANU20050318.185314/index.html.
Pełny tekst źródła"This CD contains the software (mostly written in Perl) used for comparative structure analysis reported in Chapter 4 (str_comp directory), and for the CTLD database system described in Chapter 2 (CTLD_DB directory)."
Defaus, Fornaguera Sira. "Glycoprobes for capture and identification of lectins from biological sources". Doctoral thesis, Universitat de Barcelona, 2014. http://hdl.handle.net/10803/277290.
Pełny tekst źródłaLa glicosilació, el procés enzimàtic que uneix sacàrids per produir glicans que s'adhereixen a proteïnes, lípids o altres molècules biològiques, és una modificació co-traduccional i post-traduccional present a la pràctica totalitat de components cel•lulars, on trobem glicoproteïnes, glicolípids i altres glicoderivats. Pel que fa específicament a les proteïnes, s’estima que més d’un 80% estan glicosilades i que aquests glicans són fonamentals en processos biològics com la senyalització cel•lular, el cicle infecciós de certs patògens, les respostes inflamatòria i immune, la fertilització, etc. En els últims anys s’ha avançat substancialment en el coneixement bàsic de la funció de determinats epítops o cadenes glicosídiques concretes. No obstant, es desconeixen les funcions de moltes altres estructures glicosídiques. D’altra banda, també existeix un cert desconeixement sobre les molècules que reconeixen els carbohidrats, les lectines “lectores del codi glicòmic”. Aquestes proteïnes es caracteritzen per reconèixer i unir-se de forma reversible i específica a certs monosacàrids o epítops oligosacàrids donant lloc a interaccions similars a les reaccions antigen-anticòs o enzim-substrat. El paper de les lectines en processos com l'aglutinació i la definició de grups sanguinis, la inflamació (selectines), la mielinització del teixit nerviós, la progressió tumoral, etc., demostra la ubiqüitat i diversitat d'activitats en què es veuen implicades aquestes proteïnes. Per això, disposar d'una eina ràpida i fiable per al seu aïllament i identificació, previ a l'estudi de les seves interaccions amb polisacàrids, constitueix un objectiu de màxim interès en l'actual investigació biomèdica. En la present Tesi Doctoral, es descriuen dues aproximacions complementàries mitjançant les quals es poden caracteritzar les interaccions lectina-carbohidrat amb gran sensibilitat, poca mostra i sense la necessitat de cap marcatge. En la tècnica de ressonància de plasmó superficial (SPR), el sucre és immobilitzat sobre una superfície a través d'un mòdul peptídic, la qual cosa permet (1) capturar la lectina, (2) caracteritzar la seva interacció mitjançant paràmetres cinètics i termodinàmics i (3) identificar posteriorment la proteïna mitjançant espectrometria de masses. Complementàriament, la tècnica CREDEX-MS, basada en l'excisió proteolítica del complex proteïna-sucre i posterior anàlisi per espectrometria de masses, ens permet identificar els pèptids que formen part del domini d'unió al sucre.
Tajra, FÃbio Solon. "Biotechnological potential of the red marine alga Hypnea musciformis (HML) applied to Dentistry". Universidade Federal do CearÃ, 2010. http://www.teses.ufc.br/tde_busca/arquivo.php?codArquivo=5285.
Pełny tekst źródłaDental Caries is an infectious disease naturally multi-factorial that begins in the childhood, able to spread its intensity and prevalence according to the mouth ambient conditions of each host. The risk of the patients to develop, in some time of their lives, carious lesions can be measured through a complex analysis using health determinant factors and other specific factors of the health-disease process. Since the dental caries forward diagnostic as well as the review of the auxiliary determinant factors of the dental caries danger evaluation, the treatment becomes simpler, less invasive and costing lower. It has been quite broad-casted lately the use of lectins to diagnose and to prevent some diseases. When dealing with dental caries disease, through many years this strategy has been explored still inceptively. Looking through this perspective, the goal of this trial was to investigate the biotechnological potential of the red marine alga Hypnea musciformis (HML) applied to Dentistry. It is about an explanatory-experimental study of quantitative approach through the health questionnaire and data statistics analysis. The research makes reference to some comparative analysis of salivary tests and the cariogenic activity in twelve year-old students and it searches to evaluate the biotechnological potentialities of the lectin from the red marine alga Hypnea musciformis (HML) in the diagnose and prevention of the dental caries. The concentration area of this trial was the macromolecules/ lectin biotechnological application. In the present trial were compared obtained results through the application of the conventional clinical exams to dental caries and salivary tests of dental caries activity pointed by the CARIOGRAMÂ program in association with lab analysis using specific lectins. From this trial, it has been concluded that, about lectins as a biotechnological potential to prevention and diagnose of the dental caries, it has been noticed that there is recognition of resident glycoconjugate on pacifier saliva of high and low risk observed in immunofluorescence pattern. The results showed that the tested lectin has the capacity to intervene on the bacterial growth in a negative way, it is being distinguished statistically (p<0,01) in relation to control group of saline and BSA. However, inhibition studies of the microbial biofilms using the HML are being programmed to evaluate the using possibility of these protein as biotech input to be used for dental caries prevention. It could be said also that the obtained results by the use of HML in the antimicrobial activity were promising, showing that this lectin has presented antimicrobial activity even on very small amounts. As to the use of these protein in the biofilm formation and developing there was no characteristic pattern.
A cÃrie à uma doenÃa infecciosa de natureza multifatorial que se inicia na infÃncia, podendo aumentar de intensidade e prevalÃncia de acordo com as condiÃÃes do ambiente bucal de cada hospedeiro. O risco do paciente em desenvolver, em algum perÃodo de seu ciclo de vida, lesÃes cariosas pode ser medido atravÃs de uma anÃlise complexa que utiliza os fatores determinantes da saÃde e outros fatores especÃficos do processo saÃde-doenÃa. A partir do diagnÃstico precoce da doenÃa cÃrie, assim como anÃlise dos fatores determinantes auxiliares da avaliaÃÃo do risco de cÃrie, o tratamento torna-se mais simples, menos invasivo e de menor custo. Tem sido bastante difundida ultimamente a utilizaÃÃo de lectinas para diagnÃstico e prevenÃÃo de algumas doenÃas. Em se tratando da doenÃa cÃrie, ao longo de muitos anos, essa estratÃgia tem sido explorada ainda de forma incipiente. Nesta perspectiva, o objetivo deste trabalho foi investigar as potencialidades biotecnolÃgicas da lectina de alga marinha da espÃcie de Hypnea musciformis (HML) aplicadas à odontologia. Trata-se de um estudo explicativo-experimental de abordagem quantitativa atravÃs de questionÃrio de saÃde e anÃlises estatÃsticas de dados. A pesquisa faz referÃncia à uma anÃlise comparativa de testes salivares e atividade cariogÃnica em escolares de 12 anos de idade e a busca avaliar as potencialidades biotecnolÃgicas da lectina de alga marinha da espÃcie de Hypnea musciformis (HML) no diagnÃstico e prevenÃÃo de cÃrie dentÃria. A Ãrea de concentraÃÃo deste estudo MacromolÃtulas / AplicaÃÃo BiotecnolÃgica de Lectinas. No presente estudo, foram comparados os resultados obtidos atravÃs da aplicaÃÃo de exame clÃnico convencional para cÃrie e testes salivares de atividade cariogÃnica apontados pelo programa CARIOGRAM em associaÃÃo com anÃlises laboratoriais com a utilizaÃÃo de lectinas especÃficas. A partir deste estudo, concluiu-se que, em se tratando do uso de lectinas como potencial biotecnolÃgico para a prevenÃÃo e diagnÃstico de cÃrie dentÃria, percebeu-se que hà reconhecimento de glicoconjugados presentes na saliva de pacientes de alto e baixo risco observados em padrÃo de fluorescÃncia. Os resultados demonstram que a lectina testada tem a capacidade de interferir no crescimento bacteriano de forma negativa, sendo estatisticamente significante (p<0,01) em relaÃÃo aos grupos controle de salina e BSA. Entretanto estudos de inibiÃÃo da formaÃÃo de biofilmes microbianos utilizando a HML estÃo sendo programados, para assim avaliar a possibilidade de utilizaÃÃo dessas proteÃnas como insumos biotecnolÃgicos a ser utilizados para prevenÃÃo da cÃrie dentÃria. Pode-se ainda afirmar que os resultados obtidos pelo uso de HML na atividade antibacteriana foram promissores, mostrando que esta lectina apresentou atividade antibacteriana mesmo em quantidades muito pequenas. Quanto ao uso destas proteÃnas na formaÃÃo e desenvolvimento de biofilmes, nÃo houve padrÃo caracterÃstico.
Correia, Jorge Luis Almeida. "PurificaÃÃo, caracterizaÃÃo parcial e potencialidade biotecnolÃgica de trÃs lectinas de sementes de espÃcies de Leguminosae da subtribo Diocleinae". Universidade Federal do CearÃ, 2015. http://www.teses.ufc.br/tde_busca/arquivo.php?codArquivo=17238.
Pełny tekst źródłaFundaÃÃo Cearense de Apoio ao Desenvolvimento Cientifico e TecnolÃgico
Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico
Leguminosae is recognized by the large amount of isolated and characterized lectins, especially seeds. In this group stand out proteins extracted from species belonging to subtribe Diocleinae, the number of studies in different areas of knowledge. Lectins can be defined as proteins or glycoproteins which are not originated from a body's immune response and have the ability to recognize and bind reversibly to mono or oligosaccharides particular without, however, altering their chemical structures. Different works in our group are structurally haracterizing these proteins as well as elucidating some possible biotechnological applications for these molecules. In this sense, the objective of this study was to isolate and characterize lectins of different species of Leguminosae of Diocleinae subtribe and test them for toxicity to Artemia sp. naupilos, The effect on the smooth muscle of blood vessels and the detention lectin matrix agarose previously activated with cyanogenic bromide (CNBr). Were isolated and characterized the lectin Dioclea sclerocarpa, Dioclea lasiocarpa and Dioclea lasiophylla. Were also made circular dichroism studies on lectin Dioclea sclerocarpa and Dioclea lasiocarpa. The lectin Dioclea lasiophylla was tested against Artemia sp. order to assess their toxicity and was also immobilized on agarose matrix. We evaluated the effect of lectin Dioclea lasiocarpa in the smooth muscle of blood vessels. The knowledge gained from the three scientific articles published in this thesis is a major breakthrough in this promising field of study that has been continuously growing for biotechnological applications.
A famÃlia Leguminosae à reconhecida pela grande quantidade de lectinas isoladas e caracterizadas, especialmente de sementes. Neste grupo se destacam as proteÃnas extraÃdas de espÃcies pertencentes a subtribo Diocleinae, pela quantidade de estudos em diferentes Ãreas do conhecimento. Lectinas podem ser definidas como proteÃnas ou glicoproteÃnas que nÃo sÃo originadas a partir de uma resposta imunolÃgica do organismo e possuem a capacidade de reconhecer e se ligar reversivelmente a mono ou oligossacarÃdeos especÃficos sem, no entanto, alterar suas estruturas quÃmicas. Diferentes trabalhos no nosso grupo vÃm caracterizando estruturalmente essas proteÃnas, bem como elucidando algumas possÃveis aplicaÃÃes biotecnolÃgicas para essas molÃculas. Neste sentido, o objetivo deste trabalho foi isolar e caracterizar lectinas de diferentes espÃcies de Leguminosae da subtribo Diocleinae e testa-las com relaÃÃo à toxicidade para naupilos de Artemia sp., o efeito na musculatura lisa de vasos sanguÃneos e a imobilizaÃÃo de lectina em matriz de agarose previamente ativada com brometo cianogÃnico (CNBr). Foram isoladas e caracterizadas as lectinas de Dioclea sclerocarpa, Dioclea lasiocarpa e Dioclea lasiophylla. TambÃm foram feitos estudos de dicroÃsmo circular na lectina de Dioclea sclerocarpa e Dioclea lasiocarpa. A lectina de Dioclea lasiophylla foi testada contra Artemia sp.de forma a avaliar sua toxicidade e tambÃm foi imobilizada em matriz de agarose. Foi avaliado o efeito da lectina de Dioclea lasiocarpa na musculatura lisa de vasos sanguineos. O conhecimento acumulado a partir dos trÃs artigos cientÃficos publicados nesta tese constitui um grande avanÃo no neste campo promissor de estudo que vem em contÃnuo crescimento para aplicaÃÃo biotecnolÃgica.
Sun, Yedi. "Novel Functionalized Lectins Engineered by Affinity-Guided DMAP Chemistry". 京都大学 (Kyoto University), 2013. http://hdl.handle.net/2433/174964.
Pełny tekst źródłaGemma, Emiliano. "Synthesis of Oligosaccharides for Interaction Studies with Various Lectins". Doctoral thesis, Stockholm : Department of Organic Chemistry, Stockholm University, 2005. http://urn.kb.se/resolve?urn=urn:nbn:se:su:diva-459.
Pełny tekst źródłaBanchonglikitkul, Chuleratana. "Lectins in drug delivery to the eye and mouth". Thesis, University of Portsmouth, 2000. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.323275.
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